The D-/L-Lactic Acid (D-/L-Lactate) (Rapid) test kit is used for the rapid and specific concurrent measurement and analysis of L-lactic acid (L-lactate) and D-lactic acid (D-lactate) in beverages, meat, dairy and food products.

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Grape and wine analysis: Oenologists to exploit advanced test kits.

It is without doubt that testing plays a pivotal role throughout the whole of the vinification process. To produce the best possible quality wine and to minimise process problems such as “stuck” fermentation or troublesome infections, it is now recognised that if possible testing should begin prior to harvesting of the grapes and continue through to bottling. Traditional methods of wine analysis are often expensive, time consuming, require either elaborate equipment or specialist expertise and frequently lack accuracy. However, enzymatic bio-analysis enables the accurate measurement of the vast majority of analytes of interest to the wine maker, using just one piece of apparatus, the spectrophotometer (see previous issue No. 116 for a detailed technical review). Grape juice and wine are amenable to enzymatic testing as being liquids they are homogenous, easy to manipulate, and can generally be analysed without any sample preparation.

Many of the enzymatic test kits are official methods of prestigious organisations such as the Association of Official Analytical Chemicals (AOAC) and the American Association of Cereal Chemists (AACC) in response to the interest from oenologists. Megazyme decided to use its long history of enzymatic bio-analysis to make a significant contribution to the wine industry, by the development of a range of advanced enzymatic test kits. This task has now been successfully completed through the strategic and comprehensive process of identifying limitations of existing enzymatic bio-analysis test kits where they occurred, and then using advanced techniques, such as molecular biology (photo 1), to rapidly overcome them. Novel test kits have also been developed for analytes of emerging interest to the oenologist, such as yeast available nitrogen (YAN; see pages 2-3 of issue 117 article), or where previously enzymes were simply either not available, or were too expensive to employ, such as for D-mannitol analysis.

Microbial and physicochemical succession in fermented sausages produced with bacteriocinogenic culture of Lactobacillus sake and semi-purified bacteriocin mesenterocin Y.

The inﬂuence of the bacteriocinogenic culture Lactobacillus sakei (105/g) and semi-puriﬁed bacteriocin mesenterocin Y (2560 AU/kg) on the safety and quality of traditional Croatian fermented sausages was investigated. The addition of Lb. sakei and/or mesenterocin Y reduced microbial counts (P < 0.05) in the ﬁnal products. After 28 days of ripening, coagulase-negative cocci decreased 1.5–2.0 log, yeasts 1.2–1.4 log and enterococci 1.7–2.7 log. In the case of the addition of Lb. sakei, the lactic acid bacteria count was signiﬁcantly (P < 0.05) higher at day 7 of ripening, and was accompanied by a lower pH and a higher amount of lactic acid (P < 0.05). In the ﬁnal product the amount of acetic acid was signiﬁcantly lower. More intensive proteolysis and an increase in ammonia content were found at the beginning of fermentation, and in the second phase of ripening in the control samples, respectively. The free fatty acid concentration was signiﬁcantly lower during the entire ripening process compared to the control (P < 0.05). Semi-puriﬁed mesenterocin Y did not aﬀect the sensory properties of the sausages, whilst the addition of Lb. sakei enhanced them.

Effects of yeast extract and different amino acids on the dynamics of some components in cabbage juice during fermentation with Bifidobacterium lactis BB-12.

Various amino acids and the yeast extract, in amounts of 0.1%(w/v), were separately tested for their influence on the analytical parameters of lactic acid fermentation of cabbage juice with Bifidobacterium animalis subsp. lactis BB-12. Compared with the control, cysteine supplementation led to a decrease of the time to reach pH 5.0 of 6 times and an increase of lactic acid productivity of 1.22 times. After 48 h the ascorbic acid content was by 360.73% higher, the fermented cabbage juices being assigned into a distinct group applying both factor analysis (FA) and cluster analysis (CA). Tryptophan contributed to better values for lactic and acetic acid yield, while lysine and yeast extract especially for acetic acid yield. Valine and leucine were not able to improve the fermentation progress, estimated through the analyzed variables. This work would provide some helpful information for the development of various lacto-fermented vegetable juices using probiotic bacteria.

Heterofermentative process in dry fermented sausages-a case report.

In certain circumstances the fermentation process in dry fermented sausages converts to heterofermentation pathway leading to acetic acid and carbon dioxide beside lactic acid. The study describes two cases of undesirable heterofermentation in dry sausages from two different producers. In the sausage samples (n = 7) the pH value and the content of lactic and acetic acids were measured. Microbial analysis focused on quantitative and qualitative detection of lactic acid bacteria. The acetic acid content varied from 24.28 to 67.41 µmol•g-1 dry matter, in the case of samples from the second producer the content of acetic acid (48.45 to 67.41 µmol•g-1 dry matter) was higher than the lactic acid content (20.98 to 29.02 µmol•g-1 dry matter). The lactobacilli strains from the sausages were assigned to the corresponding species by Matrix-Assisted Laser Desorption-Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) and classified to three groups according to the sugar fermentation pattern (obligately homofermentative, facultatively heterofermentative and obligately heterofermentative) and they caused the heterofermentation process in the samples of dry fermented sausages. The description of the case of heterofermentation process in dry sausages is unique and there is little information about this topic.

BACKGROUND: Safflower (Carthamus tinctorius L.), usually grown as a source of oil crop, can be used as fodder either for hay or ensiling purposes, particularly in semi-arid regions.
RESULTS: A 2-year trial was conducted in southern Italy to evaluate the production and forage quality of safflower biomass cv. Centennial, harvested at three different stages: 1, at complete appearance of primary buds (PB); 2, at complete appearance of secondary and tertiary buds (STB); and 3, at 25% of flowering stage (FS). For each stage of growth, 50% of the biomass was ensiled in 4 L glass jars without and with inoculation (Lactobacillus plantarum, LAB), and the other 50% was field wilted for 24 h before ensiling. Dry matter (DM) content and yield (DMY), pH, buffering capacity (BC) and water soluble carbohydrates (WSC) were determined on fresh forage. On safflower silages were also evaluated ammonia-N, crude protein (CP), fibre fractions, fat, lactic and acetic acids, Ca and P, and gas losses. DMY ranged from 4.5 t ha-1 (PB harvesting) to 11.6 t ha-1 (FS harvesting). DM content varied from 129 g kg-1 (PB not wilted) to 630 g kg-1 (FS wilted). The WSC in forage before ensiling with not wilting ranged from 128 (PB stage) to 105 and 100 g kg-1 DM at STB and FS stages, respectively. The wilted safflower forage showed a lower WSC compared to wilted forage. The high sugar substrate allowed lactic acid fermentation and a good conservation quality in all the harvesting stages. Silages quality was strongly influenced by the treatment performed. Wilting practice increased DM, pH and NDF contents but reduced lactic acid, acetic acid and NH3-N values. Inoculation reduced DM, pH and NDF contents, but increased lactic and acetic acids, CP and ash.
CONCLUSION: As result, wilting the forage for 1 day was very effective in the early harvesting stage because this practice significantly increased DM, reducing on the same time the intensive fermentation and proteolysis processes of silage. When harvesting is performed at the beginning of the flowering stage wilting is not necessary.

Modelling the Effect of Different Substrates and Temperature on the Growth and Lactic Acid Production by Lactobacillus amylovorus DSM 20531T in Batch Process.

Amylolytic lactic acid bacterium Lactobacillus amylovorus DSM 20531T utilised glucose, sucrose and starch as a sole carbon and energy source. The three substrates were completely depleted from MRS medium during batch cultivations carried out in a laboratory scale stirred tank bioreactor at constant temperature (40°C) and pH value (5.5). Under the tested conditions, the bacterium was capable of conducting simultaneously starch hydrolysis and fermentation. A mixture of two stereoisomers, D-(-)- and L-(+)-lactic acid, was produced in all cases by highly efficient homofermentative bioprocess with 0.93 to 1 g of lactate produced per g of total (consumed) substrate. The effect of temperature on the kinetics of cell growth and lactic acid production by the amylolytic strain in the starch-containing medium was also investigated. Efficient simultaneous saccharification and fermentation (SSF) was obtained at 35, 40 and 45°C with completely degraded complex carbohydrate in 8 to 12 h and the product yield coefficient in the range from 0.91 to 0.93 g/g. Maximum values for substrate consumption rate (0.89 h-1), maximum specific growth rate (0.87 h-1), product formation rate (2.01 h-1), and productivity of lactic acid (1.45 g/(Uh)) were obtained at 45°C, while maximum biomass concentration (4.38 g/L) was attained at 40°C. The ratio of the two stereoisomeric forms of produced lactic acid was strongly affected by the temperature. Unstructured kinetic model was used to describe the consumption of the three substrates, bacterial biomass formation and lactic acid production by L. amylovorus DSM 20531T. The dependence of biokinetic parameters on temperature was described by cardinal temperature model. The applied models successfully predicted all experimental data.

Lactose fermentation by Kombucha – a process to obtain new milk–based beverages.

This paper focuses on fermentation of lactose from a model system (black tea) and from two types of milk (0.9% w/w and 2.2% w/w of fat) by application of Kombucha. Quantities of the applied Kombucha starter were 10% v/v and 15% v/v. All fermentations were performed at 42°C. The process to achieve a desirable pH=4.5 was slower in the model system (16 h) than in milks (9 - 10 h). Regarding starter quantity, 10% v/v proved the optimal. Regarding types of milk, higher fat content guarantees shorter fermentation and higher yield of metabolites. Utilization of lactose was found at a level of ≈20% and ≈30% in milks with 0.9% w/w and 2.2% w/w of fat, respectively. This was correlated with an appearance of intermediates and/or products. Glucose underwent further transformations almost entirely, while galactose showed much lower reactivity. Seven to twelve times higher contents of lactic acid were found compared to acetic acid. Milk-based beverage from the reduced fat sample, inoculated with 10% v/v of Kombucha starter, has the best physical characteristics (syneresis and water holding capacity). It also developed a good texture (especially cohesiveness and index of viscosity). Milk lactose fermentation was a process that could have been used for obtaining new milk-based products.

A lactic acid bacterium with high tolerance of temperature and lignocellulose derived inhibitor was isolated and characterized as Pediococcus acidilactici DQ2. The strain used in the simultaneous saccharification and fermentation (SSF) for high titer lactic acid production at the high solids loading of corn stover. Corn stover was pretreated using the dry sulphuric acid pretreatment, followed by a biological detoxification to remove the inhibitors produced in the pretreatment. The bioreactor with a novel helical impeller was used to the SSF operation of the pretreated and biodetoxified corn stover. The results show that a typical SSF operation at 48°C, pH 5.5, and near 30% (w/w) solids loading in both 5 and 50 L bioreactors was demonstrated. The lactic acid titer, yield, and productivity reached 101.9 g/L, 77.2%, and 1.06 g/L/h, respectively. The result provided a practical process option for cellulosic lactic acid production using virgin agriculture lignocellulose residues.

Biohydrogen and methane production from cheese whey in a two-stage anaerobic process.

The aim of the present study was to investigate the potential of hydrogen and subsequent methane production from raw cheese whey at 35°C. The fermentative hydrogen production process from raw cheese whey was conducted in a continuous-type stirred tank bioreactor, operated at low hydraulic retention time (HRT; 24 h). In this stage, the carbohydrates contained in cheese whey are fermented to a mixture of acids and a gaseous mixture rich in hydrogen. The continuous fermentative hydrogen production was sustained by the indigenous microflora already contained in the raw cheese whey because the bioreactor was not seeded with any source of inoculum. At a HRT of 24 h, the hydrogen production rate was 7.53 L of H2/day, while the yield of hydrogen produced was 0.041 m3 of H2/kg of chemical oxygen demand (COD) added or 2.49 L of H2/L of cheese whey. The mixed liquor from this stage was further digested to biogas in a periodic anaerobic baffled reactor (PABR), a baffled-type bioreactor. The PABR was operated at HRTs of 20, 10, and 4.4 days. The highest biogas and methane production rates were 105.9 L of biogas/day and 75.6 L of CH4/day, respectively, and were obtained at an HRT of 4.4 days. During this stage, COD reduction reached 94%, obtained at an HRT of 4.4 days. Furthermore, the methane potential of the raw cheese whey was assessed by conducting a biochemical methane potential test. It was estimated to be 0.31 m3 of CH4 /kg of COD added or 17.9 L of CH4/L of cheese whey. This work demonstrated that biohydrogen production from cheese whey can be very efficiently coupled with methane production in a subsequent step, exploiting the gaseous biofuel potential of this wastewater type.

Production of functional Ricotta Cheese.

In this work, the suitability of Ricotta cheese as a food carrier for functional ingredients was evaluated. The probiotic strain Lactobacillus paracasei subsp. paracasei F19, inoculated at a concentration of 109 cfu/serving size, maintained high counts during the cold storage of Ricotta cheese (7 days at 5°C), without altering the nutritional and sensorial properties of Ricotta samples. Similarly, the addition of 3% inulin did not significantly change the sensory profile of the cheese, whereas the addition of chestnut flour lowered the perceived sensory characteristics. The synbiotic formulation (with 3% inulin and 109 cfu/serving size of Lb. paracasei subsp. paracasei F19) altered the Ricotta sensorial characteristics, mainly for an excessive acidification.

Responses in digestion, rumen fermentation and microbial populations to inhibition of methane formation by a halogenated methane analogue.

The effects of the anti-methanogenic compound, bromochloromethane (BCM), on rumen microbial fermentation and ecology were examined in vivo. Japanese goats were fed a diet of 50% Timothy grass and 50% concentrate and then sequentially adapted to low, mid and high doses of BCM. The goats were placed into the respiration chambers for analysis of rumen microbial function and methane and H2 production. The levels of methane production were reduced by 5, 71 and 91%, and H2 production was estimated at 545, 2941 and 3496 mmol/head per d, in response to low, mid and high doses of BCM, respectively, with no effect on maintenance feed intake and digestibility. Real-time PCR quantification of microbial groups showed a significant decrease relative to controls in abundance of methanogens and rumen fungi, whereas there were increases in Prevotella spp. and Fibrobacter succinogenes, a decrease in Ruminococcus albus and R. flavefaciens was unchanged. The numbers of protozoa were also unaffected. Denaturing gradient gel electrophoresis and quantitative PCR analysis revealed that several Prevotella spp. were the bacteria that increased most in response to BCM treatment. It is concluded that the methane-inhibited rumen adapts to high hydrogen levels by shifting fermentation to propionate via Prevotella spp., but the majority of metabolic hydrogen is expelled as H2 gas.

The effect of transglutaminase on rheology and texture of fermented milk products.

The aim of this study was to investigate the effect of transglutaminase (TG) addition on rheological properties, textural characteristics and microstructure of fermented milk products manufactured by different starters (probiotics and kombucha inoculum). Rheological analysis revealed that all manufactured fermented milk products had higher storage modulus than loss modulus and exhibited thixotropic and a typical shear thinning behavior. The addition of TG in milk increased approximately 10.5% hysteresis loop area, 39% firmness and 48% consistency in sample produced with probiotic starter and had more firm and stable gel structure than kombucha fermented milk products. The scanning electron microscopy micrographs showed that casein matrix of fermented milk products containing TG is continuous and uninterrupted except for void spaces occupied by milk serum and starter culture cell.